Methods and formulations for control of pests

ABSTRACT

The subject invention concerns novel biopesticides and their use to control cockroaches, carpenter ants, and pharaoh ants. Specifically, highly virulent isolates of  Beauveria bassiana  in an agricultural composition, can be used to effectively control these pests. Exemplified are  Beauveria bassiana  No. 447, ATCC 20872, and  Beauveria bassiana  SP111, ATCC 74038. Also described are unique formulations which are highly effective for delivering biocontrol agents to target pests. By using these novel compositions, target pests can be controlled without the environmental and public safety hazards presented by chemical control agents.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a continuation of application Ser. No. 08/210,965, filed Mar.21, 1994.

This application is a continuation of application Ser. No. 08/210,965,filed Mar. 21, 1994 now U.S. Pat. No. 5,683,689; which is acontinuation-in-part of application Ser. No. 07/999,185, filed Dec. 30,1992, now abandoned, which is a continuation of application Ser. No.07/687,362, filed Apr. 18, 1991, now abandoned. Application Ser. No.08/210,965 is also a continuation-in-part of application Ser. No.07/999,186, filed Dec. 30, 1992, now abandoned, which is a continuationof application Ser. No. 07/687,361, filed Apr. 18, 1991, now abandoned.

BACKGROUND OF THE INVENTION

The development of biological control agents as alternatives to chemicalinsecticides for the control of important pest species is a subject ofincreasing interest. Concerns for the environment and exposure of man toharmful substances in air, food and water have stimulated legislationand restrictions regarding the use of chemical pesticides, particularlyfor pests found in the urban environment. Control of insect pests inurban areas is highly desirable but exposure to chemical pesticides inthe household and from lawns and gardens is of great concern to thepublic. If given a choice, most people would use a non-toxic biologicalcontrol rather than a toxic chemical to control insects in the urbanenvironment. The problem is that very few biological alternatives tochemical insecticides are available for purchase and use by the public.

For most insect pests that need to be controlled in the urbanenvironment (ants, roaches, termites, fleas, wasps, etc.) there is nobiological agent available for purchase as a product.

Cockroaches are serious economic pests in urban areas. Becausecockroaches are so closely associated with humans and commonly feed ondecaying food, crumbs, or scraps, and frequent unsanitary areas such assewage systems and septic tanks, their presence leads to suspicion of athreat to human health. Pathogenic organisms have been isolated fromcockroaches collected in domestic or peridomestic environments; however,the role of cockroaches as vectors of pathogens is controversial. Unlikemany blood-feeding arthropods whose feeding behavior results in thedirect transmission of pathogens to humans, cockroaches have thepotential to transmit pathogens indirectly via contamination of foods orutensils used to prepare food. It has been demonstrated that cockroachesacquire pathogenic bacteria simply by walking over cultures and showedthat these pathogens are subsequently transferred to foodstuffs via thenormal foraging behavior of the infested cockroaches. Aside frombacterially caused food poisoning and diseases such as typhoid anddysentery, many other human illnesses and diseases associated withmicroorganisms isolated from cockroaches have been reported. Theseinclude paralytic polio, giardiasis, otomycosis, pneumomycosis, andvarious worms such as hookworm and tapeworm.

Besides the possible role of cockroaches as vectors of pathogenicmicroorganisms, the mere presence of these insects is known tocontribute to human morbidity in other ways. Perhaps the most insidiousaspect is the psychological impact of these pests in terms of theanxiety and stress related to infestation, which in some instances cantake on pathologic dimensions. Further, defensive secretions amongcockroach species may cause burning sensations, vertigo, or nausea inindividuals who come into contact with the insects.

Current cockroach control methods in buildings include preventative andcorrective approaches. Preventative measures emphasize sanitation toeliminate harborages and food sources, sealing off access routes, andthe creation of inhospitable environments by the application of boricacid or sorptive dusts in wall voids during construction (Ebeling, W.[1971] Ann. Rev. Entomol 16:123-158; Ebeling, W. [1978] UrbanEntomology, Berkeley: Univ. Calif. Div. Agric. Sci. 695 pp.). Howeverthe implementation of these measures is difficult and thus limits theireffectiveness (Thoms, E. M., W. H. Robinson [1987] J. Econ. Entomol.80:131-135). Corrective measures used to suppress establishedinfestations emphasize the use of insecticide applications. A commonlyused technique is to spray insecticides with long residual activity inareas frequented by cockroaches at fixed time intervals (Schal, C., R.L. Hamilton [1990] Ann. Rev. Entomol. 35:521-551). Despite short termsuppression of cockroach populations, toxic residues and the developmentof insecticide resistance (Cochran, D. G. [1989] J. Econ. Entomol.82:336-341) make total reliance on this technique undesirable.Alternative corrective measures such as the placement of toxic baittraps may provide sufficient control under proper conditions (Thoms &Robinson [1987], supra).

The use of natural enemies for the biological control of cockroaches hasbeen examined to varying degrees. Although traps using biocontrol agentshave been proposed, these traps are only as good as the biocontrol agentused. U.S. Pat. Nos. 5,057,315 and 5,057,316. Field releases ofparasitoids of the American and brown banded cockroaches resulted inrates of parasitism as high as 95% and has generated some optimism fortheir potential utilization (Coler, R. R., Van Driesche, R. G.,Elkinton, J. S. [1984] Environ. Entomol. 13:603-606; Hagenbuch, B. E.,R. S. Patterson, P. G. Koehler [1989] J. Econ. Entomol. 82:90-94).Pathogenic yeasts isolated from laboratory cockroach colonies also havebeen suggested as possible biological control agents, but more researchis required to evaluate their potential (Archbold, E. F., M. K. Rust, D.A. Reierson [1987] J. Med. Entomol. 24:269-272; Archbold, E. F., M. K.Rust, D. A. Reierson, K. D. Atkinson [1986] Environ. Entomol.15:221-226; Verrett, J. M., K. B. Green, L. M. Gamble, F. C. Crochen[1987] J. Econ. Entomol. 80:1205-1212). Numerous other fungi, bacteria,protozoans, and nematodes have been reported to be associated withcockroaches, but their potential as biological control agents is notsignificant, or has not been fully evaluated (Roth and Willis [1960]Smithsonian Misc. Coll. Vol. 141; Tsai, Y. H., K. M. Cahill [1970] J.Parasitol. 56:375-377; Zervos, S. [1983] N.Z. J. Zool. 10:329-334;Rahmet-Afla, M., A. F. Rowley [1989] J. Invert. Path. 53:190-196). Thus,there is a significant and longfelt need for a more effective and safemeans for controlling cockroaches.

Carpenter ants, Camponotus spp., are distributed throughout NorthAmerica. Some of the more common and/or studied species include C. modocin the Pacific northwest, C clatithorax in southern California, and theC floridanus in Florida. C. pennsylvanicus, C. noveboracensis, and C.abdominalis, are found in the east (Ebeling, W. [1978] Urban Entomology,Univ. Calif.: Berkeley p. 209-213). Public concern over carpenter antshas been increasing due to the greater probability of structuralinfestations as suburban developments extend into the forest habitats ofthe ants.

Pestiferous species of carpenter ants may be considered nuisance pestsbecause of their foraging activity inside homes. More significant damageoccurs when carpenter ants extend their nests into sound wood. Nestingsites may be located in live and dead trees, sometimes resulting indamage to shade trees. Nests may also be established in walls andsupport beams of structures, or in voids within doors, walls, andfurniture. Preference for moist or decaying wood has been reported, butnesting sites are not restricted to such areas. Carpenter antpopulations develop relatively slowly with colonies of 300-2,000 workersbeing produced over a 2-year or longer period for various species. Thepresence of reproductives follows this slow development since theirproduction has been reported only from well established colonies(Hansen, L. D., R. D. Akre [1985] “Biology of carpenter ants inWashington state (Hymenoptera: Formicidae: Camponotus),” Melandelia 43:62 pp.; Pricer, J. L. [1908] Biol. Bull. 14:177-218). Despite the slowcolony growth, large colonies with satellite colonies have been found.Worker movement occurs between the main colony and the satellites, whichserve as areas for further brood development and colony expansion(Hansen and Akre [1985], supra).

Current methods for controlling structural infestations of carpenterants include sanitation of potential and current nest sites, minimizingaccess to structures (eg. preventing the contact of tree branches with astructure), and the application of insecticides to repel (perimeterspray barriers) and/or eliminate carpenter ants. The use of boric aciddust in dry, wall voids is reported to be effective for up to 20 years(Hansen and Akre, supra).

Recommendations for the chemical control of established structuralinfestations in the home are often accompanied with warnings of possiblehazards to the applicator as well as children and pets. Alternativecontrol methods such as effective biological control agents have notbeen found (Akre, R. D., L. D. Hansen, A. L. Antonelli [1989] Ext. Bull.Washington State Univ. Coop. Ext. Serv. 1989 rev. no. EB 0818, 6 pp.). Aneed clearly exists for a safe, effective biological control agent forcarpenter ants.

Pharaoh ants, Monomorium pharaonis, have been described as “. . . themost persistent and difficult of all our house-infesting ants to controlor eradicate” (Smith, M. R. [1965] USDA-ARS Tech. Bull. No. 1326, 105pp.). It is a tropical species which has extended its range to moretemperate regions by establishing colonies in heated buildings. Pharaohants frequently infests buildings where food is prepared, and have beenfound to carry pathogenic organisms (Beatson, S. H. [1972] Lancet1:425-427).

The difficulty in controlling pharaoh ants may be attributed to theirinaccessible nesting sites, rapid population growth, and dispersion ofcolonies. Their small size allows establishment of colonies in anysuitable location, including unusual places such as between books and instored clothing. With multiple queen colonies, and the warm (30° C.),humid (63-80% RH) conditions that favor pharaoh ants, large colonies candevelop rapidly. Portions of these large colonies may disperse to formnew colonies at any time, probably in response to overcrowding andunfavorable microenvironmental conditions. Unlike other ant species,pharaoh ants do not exhibit intercolony aggression. This permits theadoption of ants from other colonies and may further enhance theestablishment of new colonies and reinfestations. Pharaoh ants alsoforage for food more than 35 m from the nest without distinct trailfollowing, and thus make nests difficult to find and eradicate.

Control methods for pharaoh ants emphasize the use of insect growthregulators (IGR) or toxicants incorporated into baits. Properlyimplemented bait programs are effective, however it may take over amonth to achieve control. Insecticide applications, while fast acting,usually do not eliminate colonies, and may be unacceptable in certainareas where toxic residues are a concern. In addition, insecticideapplications are generally not compatible with bait programs. A needexists for safe and effective biological control agents for pharaohants.

A United States patent has been granted for a fungus showing highactivity against fire ants, U.S. Pat. No. 4,925,663. This isolate,designated beauveria bassiana isolate No. 447, was deposited in a publicrepository. No biological activity other than the activity against fireants had been previously reported for this isolate, nor could activityagainst other pests be inferred from the mere knowledge that the isolatewas active against fire ants. The subject invention concerns the newuses of B. bassiana No. 447.

BRIEF SUMMARY OF THE INVENTION

The subject invention concerns the use of highly virulent Beauveriabassiana isolates to control certain pests, including cockroaches,carpenter ants, fire ants, and pharaoh ants. Specifically exemplifiedherein are formulations containing B. bassiana isolates No. 447 andSP111. These isolates, advantageously, show unexpectedly high virulenceagainst certain pests, including cockroaches, carpenter ants, fire ants,and pharaoh ants, and do not produce the environmental hazardsassociated with chemical control agents. The fungal biopesticidesdescribed herein can be applied to each of these pests in any of theirnormal habitats. The fungus may be applied, for example, directly to thepests, in trays, or applied to their surroundings, or anywhere thatthese pests are a problem. The subject invention also includes mutantsof the exemplified isolates which substantially retain the highvirulence of the parent strain.

A further aspect of the subject invention concerns unique formulationswhich can be used to effectively deliver biocontrol agents to targetpests. In a preferred embodiment a biocontrol agent is delivered in aformulation which is readily foraged by the target pest and adheres tothe body of the pest. Specifically exemplified herein is a formulationwhich has been discovered to be non-repulsive to fire ants and otherpests. This discovery is quite unexpected because pests are known to berepelled by many formulations of microbial agents. The formulation ofthe subject invention is particularly advantageous because it has beenfound to be highly effective in delivering the biocontrol agent to thetarget pest. The formulation of the subject invention comprises a uniqueblend of a food source and the fungal biocontrol agent. In a preferredembodiment, a drying agent is also used. These ingredients are presentedas a dry powder.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows cumulative percent mortality of carpenter ants exposed toB. bassiana isolate No. 447.

FIG. 2 shows cumulative percent mortality of carpenter ants exposed toB. bassiana isolate SP111.

FIG. 3 shows cumulative percent mortality of pharaoh ants exposed to B.bassiana isolate No. 447.

FIG. 4 shows cumulative percent mortality of German cockroaches exposedto B. bassiana isolate No. 447.

FIG. 5 shows cumulative percent mortality of American cockroachesexposed to B. bassiana isolate No. 447.

FIG. 6 shows cumulative percent mortality of German cockroaches exposedto B. bassiana isolate SP111.

FIG. 7 shows mortality of fire ants from the fungal formulation incomparison to chemicals for commercial traps.

FIG. 8 shows a comparison of chemical baits from traps to fungusformulation for the control of pharaoh ants.

FIG. 9 shows a comparison of chemical baits from traps to fungusformulation for the control of crazy ants.

FIG. 10 shows a comparison of chemical baits from traps to fungusformulation for the control of carpenter ants.

FIG. 11 shows a comparison of field pesticides to fungus formulation forthe control of fire ants.

DETAILED DISCLOSURE OF THE INVENTION

The subject invention concerns the use of fungal biocontrol agents tocontrol certain pests. Specifically exemplified herein is the use ofBeauveria bassiana isolates No. 447 and SP111. B. bassiana SP111 is anovel isolate. A further aspect of the subject invention includesformulations which are highly effective in delivering the biocontrolagent to the target pest.

Biologically pure cultures of Beauveria bassiana No. 447 and Beauveriabassiana SP111, have been deposited in the American Type CultureCollection (ATCC), 12301 Parklawn Drive, Rockville, Md. 20852. Thedeposit information and accession numbers are as follows:

Culture Accession Number Deposit Date Beauveria bassiana No. 447 ATCC20872 Dec. 29, 1987 Beauveria bassiana SP111 ATCC 74308 Mar. 5, 1991

The subject cultures have been deposited under conditions that assurethat access to the cultures will be available during the pendency ofthis patent application to one determined by the Commissioner of Patentsand Trademarks to be entitled thereto under 37 CFR 1.14 and 35 U.S.C.122. The deposits are available as required by foreign patent laws incountries wherein counterparts of the subject application, or itsprogeny, are filed. However, it should be understood that theavailability of the deposits does not constitute a license to practicethe subject invention in derogation of patent rights granted bygovernmental action.

Further, the subject culture deposits will be stored and made availableto the public in accord with the provisions of the Budapest Treaty forthe Deposit of Microorganisms, i.e., they will be stored with all thecare necessary to keep them viable and uncontaminated for a period of atleast five years after the most recent request for the furnishing of asample of a deposit, and in any case, for a period of at least thirty(30) years after the date of deposit or for the enforceable life of anypatent which may issue disclosing the cultures. The depositoracknowledges the duty to replace the deposit(s) should the depository beunable to furnish a sample when requested, due to the condition of adeposit. All restrictions on the availability to the public of thesubject culture deposits will be irrevocably removed upon the grantingof a patent disclosing them.

The entomopathogenic fungus Beauveria bassiana is an imperfect fungus(Fungi Imperfecti) in the subdivision Deuteromycotonia. The genusBeauveria Vuill is within the Class Deuteromycetes and is distinguishedfrom other genera by having conidia that are borne singly, notcatenulate and having the fertile portion of the conidiophore zig-zag inshape and drawn out at the tip. The species Beauveria bassiana hasspherical, not ellipsoid, conidia measuring 2-3 μm by 2-2.5 μm and withconidiophores forming dense bunches.

For a biological control agent to be effective at a practical level tocontrol cockroaches, carpenter ants, and pharaoh ants, it is essentialthat the agent not only exhibit pathogenicity against these pests, butit must also be virulent. The more virulent it is, the better it is as abiocontrol agent. Though some fungal isolates have been shown to havesome pathogenicity to these pests, these isolates did not have theessential virulence to function as a biocontrol agent. There is no knownway to convert a pathogenic non-virulent fungal isolate into apathogenic virulent isolate. Thus, the discovery of the novel isolate ofthe invention accomplishes a goal which has long been sought after.

Mode of action and virulence. Like most entomogenous fungi, Beauveriabassiana initiates infection by a germinating spore (conidium) attachingto and subsequently penetrating the cuticle of the insect host.Advantageously, and unexpectedly, the claimed Beauveria bassianaisolates attach very securely to the cuticle of cockroaches and ants andare typically not removed by the insect's grooming activities. This mayaccount somewhat for the high virulence of the fungus. As the funguspenetrates the insect's cuticle, the invasive hyphae begin to enter thehost's tissues and ramify through the hemocoel. Hyphal bodies orsegments of the hyphae distribute throughout the hemocoel, filling thedying insect with mycelia. Emergence hyphae grow out through theinsect's integument and produce spores on the external surface of thehost. These spores, or conidia, are dispersed and capable of infectingnew host insects. B. bassiana spores can be dispersed within the nest bythe activities of the pests.

Formulations. The formulations of the subject invention were found to beparticularly effective for the control of fire ants and other pests. Ina preferred embodiment, the formulation comprises a dry powder havingthe fungal biocontrol agent and a food component. Preferably, theformulation further comprises a drying agent. Optionally, theformulation may also comprise an attractant. The preferred formulationis non-repellant and includes a food source so that the target pest willforage and recruit other nestmates for foraging activity. Furthermore,the formulation of the subject invention has been found toadvantageously adhere to the body of the target pest, therebyfacilitating colonization of the pest by the fungal biocontrol agent.The ability to adhere to the pest makes the formulation of the subjectinvention quite distinct from other formulations which are currentlyused to administer chemical pesticides.

In one embodiment, the formulation of the subject invention consists ofabout 25-40% peanut material, about 45-60% cornstarch, about 2-20%fungal biocontrol agent, and about 0-15% drying agent. In a specificembodiment, the formulation can comprise about 35% peanut material,about 50% cornstarch, about 5% drying agent, and about 10% fungus. Thedrying agent can be any one of many materials known to those skilled inthe art which are small particles but have a high surface area to volumeratio so as to effectively remove water or oils from the formulation tocreate a dry powder. Preferably, the drying agent can be diatomaceousearth or a synthetic calcium silicate such as Micro-Cel®.

The peanut component of the formulation is preferably prepared bygrinding roasted peanuts so as to obtain a powder. To achieve a drypowder, it is best to grind the peanuts together with the cornstarchand/or a drying agent. Preferably, the components of the formulation aresmall particles and will pass through a 60 mesh sieve. Typically, thecornstarch and drying agent will pass through a much smaller sieve, suchas at 300 mesh. Preferably, the formulation is a powder which is freeflowing and does not stick together in clumps. Food sources other thanpeanut material or cornstarch can also be used according to the subjectinvention. The choice of a food source will depend upon the particularpest which is the target for control. Also, various attractants known tothose skilled in the art can be used. These attractants can be, forexample, pheromones or various extracts.

In a preferred embodiment, the fungal pathogen is B. bassiana No. 447 orB. bassiana SP111. However, other microbes can be used as can otherbiocontrol agents. For example, Bacillus thuningiensis can also be usedwith the formulation of the subject invention.

To evaluate the control achieved using the materials and methods of thesubject invention, tests were conducted to compare the control of pestsachieved with certain commercial pesticides. As described below, thesetests demonstrated that the fungal formulations of the present inventionare highly effective in controlling pests.

Following are examples which illustrate procedures, including the bestmode, for practicing the invention. These examples should not beconstrued as limiting. All percentages are by weight and all solventmixture proportions are by volume unless otherwise noted.

Example 1 Preparation of the Fungus

The subject fungus can be produced in trays with a rice-based medium. Anisolate of fungal inoculum is used to initiate the growth of the fungusin the trays.

The initial inoculum is prepared in petri dishes. The pure spores arethen transferred into jars containing sterile white rice without skins.

The medium for the trays is prepared as follows:

1. The rice is pre-cooked for 10 minutes.

2. 750 grams of cooked rice is placed in polyethylene bags andsterilized in an autoclave at 120° C. for 30 minutes.

3. Within a laminar flow hood, one teaspoon of spores and rice from theinoculum jars is added to each bag of prepared sterile medium.

4. Each bag is closed tightly by folding and stapling the open end.

5. The bags are transferred to a sterile room with positive pressure,temperature at 25.0-27.0° C., relative humidity above 70%, and 16 hoursphotophase. This room is known as the “environment room.”

After 3 days in the environment room, bags containing mycelia areselected and their contents are transferred to plastic trays. The sizeof the trays is such that each tray will accommodate the contents of 2-3bags. The trays and their contents are left in the environment room for12-15 days. At the end of the 12-15 day period, the trays aretransferred to a room with a cool (10-20° C.) current of clean air. Thetrays are left in this room until the cool air has dried the rice andfungus mixture.

The uncontaminated trays of rice covered with fungus can be harvestedand prepared for application or storage. If the fungus will be appliedto cockroaches or ants within 1-2 weeks after production, conidia can becollected by shaking and sieving. The resulting powder contains sporesand some mycelia, and can be applied directly to target insects or usedto prepare a formulation as a liquid, powder, or bait.

If the fungus is to be stored, the mixture can be mixed with cornstarchor talc and placed into sterile plastic containers sealed tightly andstored in a refrigerator at 4° C. or in a room with a temperature rangeof 10-25° C. and no direct sunlight. The high virulence of B. bassianacan be compromised by bacterial or fungal contamination. Therefore,throughout the preparation of the fungus, great care must be taken tomaintain the sterility of all instruments and equipment.

As the following examples demonstrate, the fungus-containing product canbe applied to target pests and their nests as a liquid, powder, or putout as a baited trap for the pests to forage, become infected, and carryinoculum back to the nest.

Example 2 Spray Application

Spraying can be used for treating individual ants or cockroaches orsmall groups of these pests. A fungal suspension containing 1.0×10⁷ to1.0×10⁹ spores per milliliter of water can be sprayed on the targetpests using an airbrush or other means as an applicator.

Example 3 Powder Application

A fungal spore and mycelia mixture can be mixed with cornstarch or talcand applied to the pests' surroundings as a dry powder.

The powder is prepared as in Example 1 above. The sieved B. bassianapowder which contains the rice, spores, and mycelia is mixed withcornstarch or talc. Application of this powder to the nests or directlyto the pests can facilitate rapid and widespread fungal growth withinthe nest or on the pest.

The application can be accomplished using an pressurized air applicatorwith an attachment that distributes the mixtures into cracks andcrevices of a pest-inhabited building. During and following application,pests covered with white powder will be observed. These infected pestswill die within 1-5 days, and the spores they produce will be infectiveto other pests. There should be a marked decrease in activity within 1-3days and death should occur within 1-2 weeks following application.Active spores will remain in the surroundings at the nest site, therebyproviding inoculum to infect other roaches or ants.

Example 4 Baited Trap Application

The fungal powder can be used in a trap in which entryways are lacedwith fungal inoculum. Preferably, fungal spores are utilized. A baitattractant contained within the trap will be foraged by cockroaches orants and the foragers will become infected. These infected individualswill return to the nest contaminated and thereby introduce the fungaldisease into the nest. A vegetable oil or other liquid substance can beadded to a bait in the trap to make it more attractive to the pests.Various attractants, including pheromone compounds, are well known tothose skilled in this art. The baited traps should be placed incabinets, along baseboards, windowsills, etc. A quantity of 0.5-2.0grams of fungal mixture containing spores and mycelia should becontained in each trap. The number of traps used in an area will dependon the level of infestation.

Example 5 Treatment of Carpenter Ants with B. bassiana No. 447

Carpenter ants (Camponotus floridanus), were exposed beauveria bassianaNo. 447. Each treatment entailed exposing two groups of 50 ants each toconidia of the isolates. Ants were coated with a conidia/cornstarchmixture, by gently shaking the ants and spores together in a coveredcontainer. The control treatment consisted of cornstarch only. Ants weresubsequently held in open plastic boxes that contained a nest cell (100mm covered petri dish with the bottom dish filled with plaster that wasperiodically moistened with water) and honey water for food.

Mortality was recorded daily for 18 days beginning with the second dayafter exposure. The test was terminated after 28 days. Dead ants wereindividually held under high humidity and examined for sporulation todetermine infection rates.

Carpenter ants exposed to isolates of B. bassiana sustained over 95%mortality (FIG. 1). At least 49% of the dead ants developed sporulatingbodies of the fungi to which they were exposed, indicating that theseisolates can grow and reproduce on carpenter ants.

Example 6 Treatment of Carpenter Ants with B. bassiana SP111

Carpenter ants (Camponotus floridanus), were exposed beauveria bassianaSP111. Each treatment entailed exposing two groups of 50 ants each toconidia of the isolates. Ants were coated with a conidia/cornstarchmixture, by gently shaking the ants and spores together in a coveredcontainer. The control treatment consisted of cornstarch only. Ants weresubsequently held in open plastic boxes that contained a nest cell (100mm covered petri dish with the bottom dish filled with plaster that wasperiodically moistened with water) and honey water for food.

Mortality was recorded daily for 18 days beginning with the second dayafter exposure. The test was terminated after 28 days. Dead ants wereindividually held under high humidity and examined for sporulation todetermine infection rates.

Carpenter ants exposed to B. bassiana SP111 sustained over 75% mortality(FIG. 2). At least 49% of the dead ants developed sporulating bodies ofthe fungi they were exposed to, indicating that these isolates can growand produce spores on carpenter ants.

Example 7 Treatment of Pharaoh Ants with B. bassiana No. 447

Pharaoh ants were exposed to a mixture comprising B. bassiana No. 447conidia as the active ingredient. Three colonies of approximately100-200 ants were individually dusted with the conidia in a petri dishand allowed to crawl out into a nest cell (15×40 mm plastic petri dishwith a plaster filled base and entrance holes in the lid). Controlsconsisting of three colonies were not dusted. The ant colonies were heldseparately in larger petri dishes along with the nest cells and honeywater. Mortality was recorded daily for 25 days. Dead ants wereindividually surface sterilized and held under high humidity to the rateof infection.

Pharaoh ant exposure to B. bassiana 447 resulted in 90% mortality after8 days (FIG. 3). Furthermore, all of the dead ants were confirmed tohave fungal spores, indicating that the fungus can successfully developon pharaoh ants.

Example 8 Treatment of Cockroaches with B. bassiana No. 447

For the German cockroach, Blattella gernanica, B. bassiana No. 447 wastested for ability of its conidia to infect and kill the host. Groups of50 male cockroaches were anesthetized with CO₂ and then dusted withconidia, within a covered container (8 oz). Controls consisted of agroup of 20 cockroaches. Dusted cockroaches were transferredindividually into separate petri dishes (10×35mm) containing moistenedfilter paper. Mortality was recorded from the second day after conidiaapplication and daily thereafter. Dead cockroaches were individuallyheld in a humidity chamber for 10 days to identify sporulating fungi.

For the American cockroach, Peniplaneta amenicana, B. bassiana No. 447was applied by brushing the conidia onto anesthetized cockroaches.Cockroaches were then held in petri dishes as described above, at 26° C.

In the test with German cockroaches the fungal isolate B. bassiana No.447 (FIG. 4) caused 100% mortality after contact with spores.Sporulation of the fungus was evident on 82% of the dead cockroaches.For the exposures of the American cockroaches, the B. bassiana isolatecaused 90% or more mortality after 8 days (FIG. 5). Fungal sporulationoccurred on all of the dead American cockroaches.

Example 9 Treatment of Cockroaches with B. bassiana SP111

For the German cockroach, Blauella gennanica, B. bassiana SP111 wastested for ability of its conidia to infect and kill the host. Groups of50 male cockroaches were dusted with conidia, within a covered container(8 oz). Controls consisted of a group of 20 cockroaches. After exposureto conidia, cockroaches were anesthetized with CO₂ and transferredindividually into separate petri dishes (10×35mm) containing moistenedfilter paper. Mortality was recorded beginning on the second day afterconidia application and daily thereafter. Dead cockroaches wereindividually held in a humidity chamber for 10 days to identifysporulating fungi.

In the test with German cockroaches the fungal isolate B. bassiana SP111(FIG. 6) caused 100% mortality after contact with spores. Sporulation ofthe fungus was evident on about 82% of the dead cockroaches.

Example 10 Evaluation of B. bassiana Bait Formulations for Control ofFire Ants

Bait formulations with ground peanut material were offered to coloniesof fire ant workers in plastic boxes (≈20×12×10 cm) containing a smalldish of water and a plastic petri dish (60 mm diameter) to serve as anest cell. Colonies were established 2-4 days before the start of theexperiment to allow ants to adapt to their new environment. Theformulations (0.5 g) were offered on weigh papers (1 square inch) orsmall dishes and left in the boxes for 3-4 days. Two controls were used:a clean control which received no formulation but only water, and a baitcontrol which received the bait formulation without any fungus.Abbott-corrected mortality greater than 70% at 14 days after treatmentwas observed for the fungal isolate containing about 10% of the B.bassiana No. 447.

Example 11 Chemical Baits from Traps Compared to Fungal Formulations forthe Control of Fire Ants

The chemical baits which were compared are from: MAX ant trap, RAID anttrap, and COMBAT ant trap. The chemical baits were removed from thetraps and offered to ants on paper. The control received the sameformulation as the fungus treatment but without conidia. The fungalformulation contained peanut material and cornstarch and 10% conidia ofB. bassiana No. 447. MAX and the fungal formulations had similarmortality, although MAX caused mortality to increase much more rapidlythan the fungus as expected, since the fungus requires 3-4 days toinfect and kill the insect. As shown in FIG. 7, COMBAT and RAID wereless efficient than MAX and Beauveria bassiana.

Example 12 Chemical Baits from Traps Compared to Fungal Formulation forthe Control of Pharaoh Ants

The chemical baits which were compared are from: MAX ant trap, RAID anttrap, and COMBAT ant trap. The chemical baits were removed from thetraps and offered to ants on paper. The control received the sameformulation as the fungus treatment but without conidia. The fungalformulation contained peanut material and cornstarch and 10% conidia ofB. bassiana No. 447. The results of this experiment is shown in FIG. 8.

Example 13 Chemical Baits from Traps Compared to Fungal Formulations forthe Control of Crazy Ants

The chemical baits which were compared are from: MAX ant trap, RAID anttrap, and COMBAT ant trap. The chemical baits were removed from trapsand offered to ants on paper. The control received the same formulationas the fungus treatment but without conidia. The fungal formulationcontained peanut material and cornstarch and 10% conidia of B. bassianaNo. 447. In all experiments, the bait with Beauveria bassiana causedmortality similar or greater than that caused by chemical baits. SeeFIG. 9 for the results of one such experiment.

Example 14 Chemical Baits from Traps Compared to Fungal Formulation forControl of Carpenter Ants

The chemical baits which were compared are from: MAX ant trap, RAID anttrap, and COMBAT ant trap. The chemical baits were removed from trapsand offered to ants on paper. The control received the same formulationas the fungus treatment but without conidia. The fungal formulationcontained peanut material and cornstarch and 10% conidia of B. bassianaNo. 447. As can be seen from FIG. 10, the fungal formulation hasperformance similar to, or slightly better than, MAX and RAID baits, andonly slightly less than COMBAT. Both COMBAT and the fungus had theireffects delayed in relation to RAID and MAX, but the delay in fungaleffect is longer than that of COMBAT.

Example 15 Field Pesticides Compared to Fungal Formulations for theControl of Fire Ants

AMDRO® chemical baits were evaluated. The bait in one experiment alsohad 10% of an ACEPHATE fire ant powder insecticide. In each treatment,one-half gram of formulation was provided per arena on weighing paper.The formulation was removed after 4 days. The control received the sameformulation as the fungus treatment but without conidia. The fungalformulation contained peanut material, cornstarch, and 10% conidia of B.bassiana No. 447.

ACEPHATE (which is not normally a bait formulation) kills the antsalmost immediately. Mortality with AMDRO increases less rapidly, but byday 4, ≈80% of the population was dead. Mortality with fungus increasesat a slower rate but final mortality after 2-4 weeks is similar to thatobtained with the chemical pesticides (FIG. 11).

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication and the scope of the appended claims.

What is claimed is:
 1. A composition for the control of a cockroach orant pest wherein said composition comprises food source particles, adrying agent, and a biocontrol agent, derived from an entomapthogenicfungi wherein said composition is a dry powder, and wherein saidbiocontrol agent is selected from the group consisting of spores andconidia and said drying agent is diatomaceous earth or silica.
 2. Thecomposition, according to claim 1, which further comprises a dryingagent.
 3. The composition, according to claim 2, wherein saidcomposition comprises about 30-40% penaut material, about 45-55%cornstarch, about 10% drying agent, and about 10% spores.
 4. Thecomposition according to claim 1, wherein said food source comprisescornstarch and ground peanut.
 5. The composition according to claim 4,wherein said composition comprises about 25-40% peanut material, about45-60% cornstarch, and about 2-20% fungal spores, wherein saidcomposition further comprises a drying agent, and wherein saidcompositions comprises less than about 15% drying agent.
 6. Thecomposition according to claimn 5, wherein said composition comprisesabout 35% peanut material, about 50% cornstarch, about 5% drying agent,and about 10% fungal spores.
 7. The composition according to claim 1,wherein said biocontrol agent is a spore.
 8. The composition accordingto claim 7, wherein said spore is a Beauveria bassiana spore.
 9. Thecomposition, according to claim 8, wherein said biocontrol agent isselected from the group consisting of Beauveria bassiana No. 447, havingall of the identifying characteristics of ATCC 20872, and Beauveriabassiana SP111, having all of the identifying characteristics of ATCC74038.
 10. The composition, according to claim 9, wherein said Beauveriabassiana is isolate No. 447, having all at the identifyingcharacteristics of ATCC
 20872. 11. The composition, according to claim9, wherein said Beauveria bassiana is isolate SP111, having all of theidentifying characteristics of ATCC
 74038. 12. The composition accordingto claim 1, wherein said biocontrol agent is conidia.
 13. Thecomposition according to claim 12, wherein said conidia is a fungalconidia.
 14. The composition according to claim 13, wherein said fungalconidia is Beauveria bassiana conidia.
 15. The composition according toclaim 14, wherein said Beauveria bassiana is selected from the groupconsisting of Beauveria bassiana No. 447, having all of the identifyingcharacteristics of ATCC 20872; and Beauveria bassiana SP111, having allof the identifying characteristics of ATCC
 74038. 16. The compositionaccording to claim 15, wherein said Beauveria bassiana is isolate No.447, having all of the identifying characteristics of ATCC
 20872. 17.The composition according to claim 15, wherein said Beauveria bassianais isolate SP111, having all of the identifying characteristics of ATCC74038.
 18. The composition, according to claim 1, wherein said particlesare capable of passing through a 65 mesh sieve.
 19. The composition ofclaim 1, further comprising a pheromone or extract.
 20. The compositionof claim 1, wherein said silica is synthetic calcium silicate.
 21. Acomposition for the control of a cockroach or ant pest wherein saidcomposition comprises food sources particules, a drying agent, and abiocontrol agent, wherein said composition is a dry powder capable ofpassing through a 300 mesh sieve, and wherein said biocontrol agent isselected from the group consisting of spores and conidia and dryingagent is diatomaceous earth or silica.
 22. The composition according toclaim 21, wherein said food source comprises cornstarch and groundpeanut.
 23. The composition according to claim 21, wherein saidbiocontrol agent is a spore.
 24. The composition of claim 21, furthercomprising a pheromone or extract.
 25. The composition of claim 21,wherein said silica is synthetic calcium silicate.
 26. The compositionaccording to claim 21, wherein said biocontrol agent is conidia.
 27. Thecomposition according to claim 26, wherein said composition comprisesabout 45-60% cornstarch, about 2-20% fungal spores, about 25-40% peanutmaterial, and wherein said composition further comprises less than about15% drying agent.
 28. The composition according to claim 27, whereinsaid composition comprise about 35% peanut material, about 50%cornstarch, about 5% drying agent, and about 10% fungal spores.
 29. Thecomposition according to claim 26, wherein said fungal spores areBeauveria bassiana spores.
 30. The composition, according to claim 29,wherein said Beauveria bassiana selected from the group consisting ofBeauverian bassiana No. 447, having all of the identifyingcharacteristics of ATCC 20872, and Beauveria bassiana SP111, having allof the identifying characteristics of ATCC
 74038. 31. The composition,according to claim 30, whererein said Beauveria bassiana is isolate No.447, having all of the identifying characteristics of ATCC
 20872. 32.The composition, according to claim 30, wherein said Beauveria bassianais isolate SP111, having all of the identifying characteristics of ATCC74038.